PRMT2稳定低表达对MCF-7细胞自噬的影响及机制研究

发布时间：2018-03-21 07:57

本文选题：蛋白精氨酸甲基转移酶2(PRMT2)　切入点：自噬　出处：《南华大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:明确蛋白精氨酸甲基转移酶2(protein arginine methyltransferase 2,PRMT2)对人乳腺癌MCF-7细胞自噬的影响,并初步探讨其可能的分子机制。方法:根据MCF-7细胞中PRMT2表达水平分为PRMT2稳定低表达组及PRMT2正常水平表达组。首先,蛋白质免疫印迹法(Western Blot,WB)分别检测两组细胞中自噬相关蛋白Beclin1、LC3(light chain3,轻链3)的表达,初步探讨PRMT2低表达对细胞自噬的影响。其次,WB方法分别检测两组细胞中自噬信号通路分子mTOR、ULK1、p-AMPK、AMPK(mTOR:mammalian target of rapamycin,雷帕霉素靶蛋白;ULK1:unc-51-like kinase 1,unc-51样激酶1;AMPK:Adenosine monophosphate-activated protein kinase,单磷酸腺苷活化蛋白激酶)的表达并进行分子机制的初步研究。第三,应用免疫共沉淀方法初步研究PRMT2影响自噬的可能信号通路。结果:1.WB检测结果表示:PRMT2稳定低表达的MCF-7细胞中,自噬相关蛋白(Beclin1、LC3)表达增加。2.PRMT2稳定低表达的MCF-7细胞中,mTOR、ULK1、p-AMPK表达均上调,AMPK表达水平与普通MCF-7组无明显差异。3.免疫共沉淀结果表示:MCF-7细胞中,PRMT2与AMPK之间以及Beclin1与PRMT2及AMPK之间均存在相互作用。结论:MCF-7细胞中,PRMT2可能通过与AMPK相互作用,经AMPK/ULK1/Beclin1途径,影响细胞自噬水平。
[Abstract]:Objective: to investigate the effect of protein arginine methyltransferase 2 protein arginine methyltransferase 2 (PRMT2) on autophagy of human breast cancer MCF-7 cells. Methods: according to the level of PRMT2 expression in MCF-7 cells, it was divided into two groups: PRMT2 stable low expression group and PRMT2 normal expression group. Western blotblotWB was used to detect the expression of the autophagy associated protein Beclin1 (LC3) chalight 3 (light chain 3) in two groups. To investigate the effect of low expression of PRMT2 on autophagy. Secondly, we detected the autophagy signal pathway molecule mTOROULK1, p-AMPK1, AMPC: mammalian target of rapamycin, ULK1: unc-51-like kinase 1 unc-51 like kinase 1 AMPK1: Adenosine monophosphate-activated protein kinase1, adenosine monophosphate activated protein. Enzyme) expression and preliminary study of molecular mechanism. Third, The possible signal pathway of PRMT2 affecting autophagy was studied by immunoprecipitation method. The expression of AMPK in MCF-7 cells with stable and low expression of PRMT2 was not significantly different from that in normal MCF-7 group. 3. The immunoprecipitation results showed that the expression of Beclin1, PRMT2 and AMPK in MCF-7 cells was higher than that in normal MCF-7 cells. 3. The results of immunoprecipitation showed that the expression of AMPK in MCF-7 cells with stable and low expression of PRMT2 was not significantly different from that in normal MCF-7 cells. 3. The results of immunoprecipitation indicated that there was no significant difference between Beclin1 and AMPK in MCF-7 cells and between Beclin1 and PRMT2 and AMPK. Conclusion PRMT2 may interact with AMPK in MCF-7 cells. The level of autophagy was affected by AMPK/ULK1/Beclin1 pathway.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R737.9

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